po 237 neutralising extracellular morgana impairs breast tumour growth and migration

Introduction Morgana is a ubiquitously expressed protein with chaperone activity per se and HSP90 co-chaperone function. High Morgana expression correlates with high tumour grade, mitosis number, and lymph node positivity in different breast cancer subtypes. Several chaperones are overexpressed in a wide range of human cancers and are implicated in tumour progression. Moreover, it has become evident that also from the extracellular compartment, where surprisingly chaperones and co-chaperones are actively released by cancer and immune cells, they favour tumour progression. If, the cytoplasmic role of Morgana has been well characterised in tumorigenesis and metastasis formation of Triple-Negative Breast Cancer (TNBC), nothing is known about extracellular Morgana (eMorgana). Material and methods Conditioned medium from human and murine TNBCs cell lines were analysed for Morgana presence. To address the role of eMorgana, a Maltose Binding Protein (MBP) fused recombinant protein was produced in ClearColi BL21 and used to evaluate eMorgana role in migration, treating MDA-231 and BT-549. The identification of Morgana receptor was performed indirectly, through the inhibition of Toll-like 2 and Toll-like 4 receptors (TLR2, TLR4). The activity of extracellular Morgana was inhibited, in mice injected with the syngenic cancer cell line E0771, using the homemade blocking antibody 5B11. Results and discussions Morgana is secreted by TNBC cell lines and as for other chaperones and co-chaperones, Morgana reaches the outside with an unconventional mechanism. From the extracellular compartment Morgana is able to induce cell migration, through theTLR2 and TLR4. Blocking of eMorgana with 5B11 inhibits migration in vitro and proliferation in vivo. Different efforts are needed to understand if Morgana binds to TLRs alone or through HSP90 and the consequently downstream signalling pathway. Moreover since TLR are prevalently expressed by immune cells it is important to address the role of eMorgana in this context and the possible crosstalk between the tumour and microenvironment. Conclusion Since TNBCs have an high rate of recurrence and poor prognosis, the identification of innovative treatments is an urgent need. In this view, although many other studies are required, eMorgana in serum patients could represent a new biomarker and its targeting a possible clinical approach in breast cancers

PO-356 MicroRNA mediated regulation of morgana, a new oncosuppressor in chronic myeloid leukaemia

Introduction Morgana is a chaperone protein encoded by the CHORDC1 gene. Its deletion is embryonic lethal due to apoptosis of the cells of the inner cell mass. We recently characterised the role of Morgana in myeloid malignancies, as the haploinsufficiency of the protein in mice is able to induce a fatal and transplantable myeloproliferative disease resembling human Atypical Chronic Myeloid Leukaemia (aCML). 5 out of 5 aCML patients and 16% of Philadelphia-positive CML patients express low/undetectable levels of Morgana in their bone marrow. As we never found mutations or deletions of CHORDC1 gene, we decided to investigate if Morgana can be targeted by miRNAs. Five miRNAs are predicted to target Morgana: miR-15a/b and miR-16 sharing the same seed sequence and miR-26a/b. Material and methods HEK-293T cells were used to overexpress miRNAs predicted to target Morgana mRNA. The level of miRNAs overexpression and Morgana mRNA was assessed with qRT-PCR and Morgana protein level with Western Blot at different time points. The seed sequences for the selected miRNAs in the 3'UTR of CHORDC1 gene were than mutagenized to validate the specificity of the binding. Bioinformatic analysis were used to correlate miRNAs and Morgana expression levels in leukaemia and lymphoma. Results and discussions We demonstrated that miRNA-15a/b and miRNA-26a/b are able to bind to Morgana 3'-UTR and, in this way, mediate its mRNA deregulation leading to a reduction of Morgana, both at mRNA and protein level. We were able to highlight an anti-correlation between Morgana and miRNAs expression in haematological tumours: in particular miR-15b in Chronic Lymphocytic Leukaemia and Lymphomas, miR-15a in aCML and CML and miR-26a and miR-16 in Lymphomas. Conclusion Morgana is able to act both as proto-oncogene and as oncosuppressor depending on tissue type and levels of expression as it is frequently found both overexpressed and downregulated. Different approaches to elucidate its mechanisms of regulation failed and we believe that miRNAs are just one of them. Further investigations are needed to clirify how Morgana expression is regulated in different type of tumours

Domain Formalization for Metaphorical Reasoning

It is commonplace that cultural heritage is always discussed and analysed by using references to figures of speech. In particular, metaphors and allegories are very frequent in ancient and historical documents, painting and sculptures. It is also frequent to have some hints about the assets and their authors by comparing their contents with elements in the domain of the figures of speech to which the asset refer. In order to enable reasoning by figures of speech, we propose here a methodology able to link concepts in the domain of cultural assets with concepts in the domain of the figure of speech. We show here how reasoning in all these domains help in discovering some elements related to the cultural heritage that humans may neglect at first glance. Our approach can be useful in a variety of applications related to cultural heritage such as semantics annotations, linked data, crowd-sensing, etc. © 2019, Springer Nature Switzerland AG